RNA-seq protocol for rRNA depletion

The total RNA (1.0 μg/sample) was subjected to rRNA depletion by Ribo-Zero rRNA Removal (Human/Mouse/Rat) Kit (Illumina, San Diego, CA, United States). Strand-specific RNA libraries were prepared using TruSeq Stranded Total RNA Sample Preparation Kit (Illumina) without poly-A selection. All assays were performed according to the manufacturer’s instructions. RNA-seq was performed on the Illumina Nova6000 plate High Output Model (Illumina, San Diego, United States) (Hrdlickova et al., 2017 (link)), in a 2 × 150-bp paired-end configuration, with a total of more than 2,666 M reads per lane (at least 50 M reads per sample).

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Su S., Li M., Wu D., Cao J., Ren X., Tao Y.X, & Zang W. (2021). Gene Transcript Alterations in the Spinal Cord, Anterior Cingulate Cortex, and Amygdala in Mice Following Peripheral Nerve Injury. Frontiers in Cell and Developmental Biology, 9, 634810.

Publication 2021

Ribo-Zero rRNA Removal (Human/Mouse/Rat) Kit TruSeq Stranded Total RNA Sample Preparation Kit Nova6000 plate High Output Model

Assays Human Mouse Poly a Rna seq Rrna

Corresponding Organization : Rutgers, The State University of New Jersey

Other organizations : First Affiliated Hospital of Zhengzhou University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

RNA-seq analysis

control variables

Total RNA (1.0 μg/sample)
RRNA depletion by Ribo-Zero rRNA Removal (Human/Mouse/Rat) Kit (Illumina)
Strand-specific RNA libraries prepared using TruSeq Stranded Total RNA Sample Preparation Kit (Illumina) without poly-A selection
Assays performed according to the manufacturer's instructions
RNA-seq performed on the Illumina Nova6000 plate High Output Model in a 2 × 150-bp paired-end configuration
More than 2,666 M reads per lane (at least 50 M reads per sample)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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